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Updated: May 26, 2026
Mass Spectrometry-Guided Genome Mining as a Tool to Uncover Novel Natural Products
Published on: March 12, 2020
Genome Mining and Characterization of an Uncharacterized Monofunctional Class I Diterpene Synthase From a
Mengxiao Zhang1,2, Shuxuan Li1, Hanqi Huang3
1National Key Laboratory for the Development and Utilization of Forest Food Resources, Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University, Nanjing, China.
Abstract:
The first marine-derived fungal monofunctional class I diterpene synthase RousA was characterized by a combination of bioinformatics, phylogenetic analysis, and heterologous coexpression. RousA resides on a remote branch of phylogenetic tree, indicating it is an uncharacterized terpene cyclase. Heterologous coexpression of RousA and RousB in engineered yeast led to the production of a 5-8-5 tricyclic diterpene scaffold 1. Its absolute configuration was first established by ECD calculation. Structural modeling and site-directed mutagenesis demonstrated that several amino acid residues in nonconserved motif have a strong influence on 5-8-5 accumulation and F65 residue serves as pivotal "bidirectional functional switch" between tricyclic and bicyclic systems. A cyclization process of tricyclic product 1 exhibits a deprotonation mechanism distinct from those of known enzymes. The antimicrobial and plant immune activities were also evaluated.
